Cytogenetics on released DNA fibers.
DNA fibers can be released from cell suspensions and frozen tissue and can be used as a template for hybridization with multiple differently labeled probes. Simultaneous hybridization with 5-10 adjacent or partly overlapping probes generates a highly specific "color barcode" for individual DNA segments. Rearrangements in this barcode can be easily detected and mapped. The resolution of DNA fiber FISH is between 2 and 500kb. In mixing experiments of cell lines with different structural abnormalities, we found a sensitivity of approximately 10%. We applied DNA fiber FISH for detection of t(11;14) in mantle cell lymphoma (MCL) and immunoglobulin (Ig) class switching in hairy cell leukemia (HCL). Using a barcode for the Ig and BCL-1 loci at 14q32 and 11q13, we detected and mapped a t(11;14) breakpoint in 35 of 36 MCL. In 5 cases complex mono-allelic rearrangements at both sides of the cyclin D1 gene were identified. In 13 HCL with phenotypic evidence of Ig class switching, including 2 cases with solely IgD, fiber FISH revealed concordant Ig class switch deletions. In most cases both alleles were affected. These results indicate that DNA fiber FISH is a very powerful method to detect and map structural DNA alterations.
Vaandrager
J W
JW
Dept. of Pathology, Leiden University Medical Center, The Netherlands.
Kleiverda
J K
JK
Schuuring
E
E
Kluin-Nelemans
J C
JC
Raap
A K
AK
Kluin
P M
PM
D002880:Chromosomes, Human, Pair 11; D002883:Chromosomes, Human, Pair 14; D015342:DNA Probes; D004273:DNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D005803:Genes, Immunoglobulin; D005812:Genes, Switch; D006801:Humans; D007134:Immunoglobulin Switch Region; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007943:Leukemia, Hairy Cell / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D008228:Lymphoma, Non-Hodgkin / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D014178:Translocation, Genetic
D015342:DNA Probes; D004273:DNA, Neoplasm